Comparison of sample preparation methods for ChIP-chip assays

Henriette O'Geen, Charles M. Nicolet, Kim Blahnik, Roland Green, Peggy J. Farnham

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122 Scopus citations

Abstract

A single chromatin immunoprecipitation (ChIP) sample does not provide enough DNA for hybridization to a genomic tiling array. A commonly used technique for amplifying the DNA obtained from ChIP assays is ligation-mediated PCR (LM-PCR). However, using this amplification method, we could not identify Oct4 binding sites on genomic tiling arrays representing 1% of the human genome (ENCODE arrays). In contrast, hybridization of a pool of 10 ChIP samples to the arrays produced reproducible binding patterns and low background signals. However, the pooling method would greatly increase the number of ChIP reactions needed to analyze the entire human genome. Therefore, we have adapted the GenomePlex® whole genome amplification (WGA) method for use in ChIP-chip assays; detailed ChIP and amplification protocols used for these analyses are provided as supplementary material. When applied to ENCODE arrays, the products prepared using this new method resulted in an Oct4 binding pattern similar to that from the pooled Oct4 ChIP samples. Importantly, the signal-to-noise ratio using the GenomePlex WGA method is superior to the LM-PCR amplification method.

Original languageEnglish (US)
Pages (from-to)577-580
Number of pages4
JournalBioTechniques
Volume41
Issue number5
DOIs
StatePublished - Nov 2006

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ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

O'Geen, H., Nicolet, C. M., Blahnik, K., Green, R., & Farnham, P. J. (2006). Comparison of sample preparation methods for ChIP-chip assays. BioTechniques, 41(5), 577-580. https://doi.org/10.2144/000112268